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Good starter CP's for TC?

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Nov 15, 2015
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For a while now, I've been wanting to do a bit of tissue culture. I'm still organizing information and supplies, but I realized I had no idea what to TC. I don't want to buy some expensive material and fail, and I don't have a very extensive plant collection. I am wondering, you jelly people, what did you first use to clone? I think my best option would to work with an African violet, and get into, say, drosera, as I become better. Thoughts? (horrible way to end).
 
Joined
Mar 11, 2010
Messages
572
For a while now, I've been wanting to do a bit of tissue culture. I'm still organizing information and supplies, but I realized I had no idea what to TC. I don't want to buy some expensive material and fail, and I don't have a very extensive plant collection. I am wondering, you jelly people, what did you first use to clone? I think my best option would to work with an African violet, and get into, say, drosera, as I become better. Thoughts? (horrible way to end).
Sweet! Another TC guy around here.

I've yet to do much explant work this time around, but I've been having good results with all kinds of seeds. I just haven't had much live tissue to work with, until recently, now that my seedlings are finally putting on some size. I also want to pick up an African Violet to practice on, just because of their status as the "starting point" for explant work. The fuzzy leaves aren't necessarily ideal for cleanliness, though, unless you grow it in a bag of under a dome for long enough for a complete new leaf to be produced and removed before it gets a chance to collect dust.

I recommend you start with seeds, as they can generally handle harsher sterilization than explants, which should lead to more initial success while you get your sterile protocol down, and cost less money than buying live rare plants to chop up. Don't expect to get it perfect in the beginning. If you do, it's a miracle, but don't be discouraged if you get lots of contamination early on.

I'd start with fast germinating seeds, and thick tissue for explants, such as...
(in rough order of ease with the top being the easiest)
-Basil seeds/stems (the seeds actually have a gell coating that forms when they are wet. I've had success with them, even without removing it.)
-Dionaea seed
-D. capensis seed
-Pygmy Drosera gemmae (super fast sprouting + explant sterilization practice)
-Dionaea flower stalks
-Dionaea leaves

Pinguicula leaves are super sensitive and fragile. It's extremely easy to kill them. Drosera leaves are good, but I recommend you grow one bagged, or under a dome until you get a new leaf, like with the African Violets. Otherwise the dew will collect up all kinds of contaminants.

Basil seed:
9C3E2A26-F52F-4827-9FDD-41310F751CC8_zps1xazsrhj.jpg

2D84AD8B-5437-4313-A2A0-E061780E3511_zpsclkuzxmv.jpg


Dionaea seed:
43CEF65E-CC85-4A0B-A626-2E515459A53E_zpsm28skca3.jpg


Drosophyllum lusitanicum seedling:
88194097-76CC-4FE3-83FA-413DBAA6F516_zpskmd0nnjq.jpg
 
Joined
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Messages
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In a Glalaxy far, far away
First of all, thank you for such a detailed reply! I think what I might do is wait until spring, since my bog will wake up, and I'm always wondering what to do with all the VFT seeds. I'm currently setting up a basic cleanbox. An old fishtank, resealed in fishtank silicone, vinyl shower curtain, and two holes I'm "welding" kitche gloves into. I can't afford a commercial growbox (I don't think many people can either), so I have to improvise on a lot of things. I guess I'm eating Paul Newman's salsa for the next couple of months:-O.
 
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First of all, thank you for such a detailed reply! I think what I might do is wait until spring, since my bog will wake up, and I'm always wondering what to do with all the VFT seeds. I'm currently setting up a basic cleanbox. An old fishtank, resealed in fishtank silicone, vinyl shower curtain, and two holes I'm "welding" kitche gloves into. I can't afford a commercial growbox (I don't think many people can either), so I have to improvise on a lot of things. I guess I'm eating Paul Newman's salsa for the next couple of months:-O.
No problem, man!

This time around, I've been working in a DIY glove box I built. I actually haven't even been using the big arm length gloves, and have just been using regular latex (powder-free) ones. I spray my arms and the gloves with IPA, and spray the air in the box as well. It seems like it keeps the air still enough in there to stop most airborne contamination, and it's much easier than using those thick yellow gloves. I feel safer in terms of sterility, with the yellow gloves, but I can work faster with the latex ones which also helps prevent contamination, so it's a trade-off.

81D15AB5-C685-498C-A18D-BA00D2988B13_zpslpwjqmi7.jpg
 
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Joined
Nov 15, 2015
Messages
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In a Glalaxy far, far away
Again, thank you for the information! I think I might copy your glove box idea, it seems quite simple. I can now save the tank for some other valuable purpose (sitting in a closet). What percentage of IPA do you use? I've heard that 70% works, but I'm trying to get as many different views as possible. Also, is their a chance of bacteria going through the lid on the bottom? I've done a small amount of mushroom culturing, but it is nothing near as hard as this.
 
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Again, thank you for the information! I think I might copy your glove box idea, it seems quite simple. I can now save the tank for some other valuable purpose (sitting in a closet). What percentage of IPA do you use? I've heard that 70% works, but I'm trying to get as many different views as possible. Also, is their a chance of bacteria going through the lid on the bottom? I've done a small amount of mushroom culturing, but it is nothing near as hard as this.
You're welcome! My glove-box isn't airtight, even with the arm gloves, but if you arent pushing and pulling on the sides too much, I don't see much contamination getting in though the bottom rim. Especially, since the whole thing is wet with IPA when I close it up. You can try some silicone maybe or some other non-porous weatherproofing strips for the edge, but I don't thinkk it's necessary. I would prefer a flat lid (the "floor" of the box), though. I work with the arm holes open lately, and haven't been getting any media surface contamination. If I get any, it originates from the plant material, so I think airborne contamination is pretty well controlled in there.

As for the IPA, I've been using 91% from Walgreens or any other pharmacy. Some only have 70%, which is supposed to be ok, but the 91% dries up faster.

I recommend picking up some 3M Micropore tape, or some other breathable, low-adhesive tape, to wrap your vessel lids with, too. That helps prevent contamination from getting in for the weeks/months after plating.
 
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Thank you for all the advice! I thin I'm going to start with basil, since I can get it anywhere for cheap, and wait for a heatwave this winter to get a few drosera. I also recenty acquired a grow rack, and I'm clearing an old cellar to make a plant/TC room. I'll try to post pic, but the only lighting is a measly incandesant bulb as old as the house.
 
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Good idea. For your TC room, try to use surfaces that can be sprayed with bleach and wiped down. That'll help you get it nice and clean before each "operation". Also, I'd recommend throwing an air purifier in there, especially if you are going to have plants nearby (which is pretty risky). Also, tape your culture vessels...seriously. If you don't, and you have plants in your house, little bugs like springtails, will find their way into the jars, and ruin them all.

Feel free to send me a pm (here or on instagram @abugsnightmare) or comment on one of my threads, if you need any help, or to discuss more TC stuff.
 
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Very interesting post.

I have actually been keen on starting my own TC setup but don't have the time or space at the moment.
So far I have just been doing research on the subject.

Let us know how your setup turns out [MENTION=11978]Masdevallia[/MENTION]
 
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Nov 15, 2015
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Very interesting post.

I have actually been keen on starting my own TC setup but don't have the time or space at the moment.
So far I have just been doing research on the subject.

Let us know how your setup turns out [MENTION=11978]Masdevallia[/MENTION]

I'm in your shoes right now. I've spent forever now researching and clearing an area for my lab. I've finally cleared an area, but still think I could do more research. You cant do enough!
 
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I'm in your shoes right now. I've spent forever now researching and clearing an area for my lab. I've finally cleared an area, but still think I could do more research. You cant do enough!

Hey [MENTION=11978]Masdevallia[/MENTION], how is you lab coming along?
 
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Hey [MENTION=11978]Masdevallia[/MENTION], how is you lab coming along?

Quite nicely. I have everything pretty much at this point. I'm just looking for a good pressure cooker that wont break the bank, or other sterilization means. Hoping to start it up when its not a winter "wonderland".




edit: Pressure cookies. delicious
 
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Joined
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Awesome!
Why exactly a pressure cooker though? Can't you simply boil the equipment in water?
Water boils at too low of a temperature to effectively sterilize all microbes. You'd be surprised how resilient these single cell organisms are.
 
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Water boils at too low of a temperature to effectively sterilize all microbes. You'd be surprised how resilient these single cell organisms are.

It is not so much just an issue of temperature; but also the increase in atmospheric pressure offered by a pressure cooker / autoclave, which ensures sterility. Find a cooker which allows for at least 15 psi setting; anything below that is worthless . . .
 
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